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J. Bacteriol. doi:10.1128/JB.01224-06
Copyright (c) 2006, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Glycosylation of Pseudomonas aeruginosa strain Pa5196 type IV pilins with mycobacterial-like -1,5 linked D-Araf oligosaccharides

Institute for Biological Sciences, National Research Council, Ottawa, ON, Faculty of Dentistry, University of Toronto, Toronto, ON and Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON

	Abstract

Pseudomonas aeruginosa is a Gram negative bacterium that uses polar type IV pili for adherence to various materials and for rapid colonization of surfaces via twitching motility. Within the P. aeruginosa species, five distinct alleles of the structural subunit PilA have been identified, varying in amino acid sequence, length and presence of post-translational modifications. In this work, a combination of mass spectrometry and NMR spectroscopy was used to identify a novel glycan modification on the pilins of the group IV strain Pa5196. Group IV pilins continued to be modified in a lipopolysaccharide (wbpM) mutant of Pa5196, showing that, unlike group I strains, the pilins of group IV are not modified with the O antigen unit of the background strain. Instead, the pilin glycan was determined to be an unusual homo-oligomer of 1, 5-linked D-arabinofuranose (D-Araf). This sugar is uncommon in prokaryotes, occurring mainly in the cell wall arabinogalactan (AG) and lipoarabinomannan (LAM) polymers of mycobacteria, including Mycobacterium tuberculosis and M. leprae. Antibodies raised against M. tuberculosis LAM specifically identified the glycosylated pilins from Pa5196, confirming the glycan is antigenically, as well as chemically, identical to those of Mycobacterium. P. aeruginosa Pa5196, a rapidly-growing strain of low virulence that expresses large amounts of glycosylated type IV pilins on its surface, represents a genetically tractable model system for elucidation of alternate pathways for biosynthesis of D-Araf and its polymerization into mycobacterial-like 1,5 linked oligosaccharides.

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